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Status |
Public on Mar 31, 2013 |
Title |
Lung-X31-12h-S3 |
Sample type |
RNA |
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Source name |
wild-type C57BL/6J mice lung tissue
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Organism |
Mus musculus |
Characteristics |
strain: C57BL/6J infection (influenza strain): X31 time (hours): 12
|
Treatment protocol |
Virus aliquots stored at -80C; quick thaw to RT and immediate dilution. Viruses diluted to 1x10^5 PFU/30ul (3.33x10^6 PFU/ml) in ice-cold 1x PBS in polystyrene tubes. Virus dilution maintained at 4C until use. Mice are all female C57BL/6J from Jackson Lab.
|
Extracted molecule |
total RNA |
Extraction protocol |
Mice were sacrificed by CO2 inhalation. Whole lungs (right side) removed and homogenized (with bead homogenizer) in 1 ml Trizol. Samples snap frozen in liquid nitrogen.
|
Label |
Cy3
|
Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 200 ng total RNA using Agilent’s Low Input Quick Amp One-Color Labeling Kit according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and
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Hybridization protocol |
600 ng of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 25ul containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions.
|
Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner using the Agilent G3-1-color scan setting for 8x60k arrays.
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Description |
Lung tissue after stimulation with either PR8, VN, X31 or Mock for 12 hours
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Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.3.1 (Agilent) using default parameters (protocol GE1_107_Sep09 and Grid: 028005_D_F_20101029).
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Submission date |
Nov 14, 2012 |
Last update date |
Mar 31, 2013 |
Contact name |
Aderem Lab |
E-mail(s) |
[email protected]
|
Organization name |
Seattle Children's Research Institute
|
Lab |
Aderem Lab
|
Street address |
307 Westlake Avenue North, Suite 500
|
City |
Seattle |
State/province |
WA |
ZIP/Postal code |
98109 |
Country |
USA |
|
|
Platform ID |
GPL10787 |
Series (1) |
GSE42285 |
Host response in the lung to influenza infection with PR8, VN or X31 strains at 12, 16 and 24 hours |
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